Innate immune activation in neonatal tracheal aspirates suggests endotoxin-driven inflammation.

Pediatric Research
Katheryn E NatheOfer Levy

Abstract

Tracheal aspirates (TAs) from critically ill neonates accumulate bacterial endotoxin and demonstrate mobilization of endotoxin-binding proteins, but the potential bioactivity of endotoxin in TAs is unknown. We characterized innate immune activation in TAs of mechanically ventilated neonates. Innate immune activation in TAs of mechanically ventilated neonates was characterized using a targeted 84-gene quantitative real-time (qRT) PCR array. Protein expression of cytokines was confirmed by multiplex assay. Expression and localization of the endotoxin-inducible antimicrobial protein Calgranulin C (S100A12) was assessed by flow cytometry. Endotoxin levels were measured in TA supernatants using the Limulus amoebocyte lysate assay. Analyses by qRT-PCR demonstrated expression of pattern recognition receptors, Toll-like receptor-nuclear factor κB and inflammasome pathways, cytokines/chemokines and their receptors, and anti-infective proteins in TA cells. Endotoxin positivity increased with postnatal age. As compared with endotoxin-negative TAs, endotoxin-positive TAs demonstrated significantly greater tumor necrosis factor (TNF), interleukin (IL)-6, IL-10, and serpin peptidase inhibitor, clade E, member 1 (SERPINE1) mRNA, and IL-10, TN...Continue Reading

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Citations

Oct 14, 2017·The Journal of Antimicrobial Chemotherapy·Marjan M HashemiPaul B Savage
Jan 17, 2014·Pediatric Research·Olaf Dammann, Alan Leviton
Nov 22, 2013·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Nicola F FletcherJane A McKeating
Nov 13, 2013·Expert Review of Clinical Immunology·Kirsten Glaser, Christian P Speer
Sep 2, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·Dietmar SpenglerMartin F Krause
Nov 13, 2020·Journal of Perinatology : Official Journal of the California Perinatal Association·Christiana N Oji-MmuoPatricia Silveyra

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Methods Mentioned

BETA
enzyme-linked immunosorbent assay
confocal microscopy
bronchoalveolar lavage
protein assay
flow
Flow Cytometry

Software Mentioned

SA Biosciences
Graphpad Prism
FloJo Flow Cytometry
QIAGEN
Slidebook

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